+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 7pbq | ||||||
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タイトル | RuvAB branch migration motor complexed to the Holliday junction - RuvB AAA+ state s0+A [t2 dataset] | ||||||
要素 |
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キーワード | HYDROLASE / DNA recombination / DNA repair / branch migration / Holliday junction / helicase | ||||||
機能・相同性 | 機能・相同性情報 Holliday junction helicase complex / Holliday junction resolvase complex / four-way junction helicase activity / four-way junction DNA binding / DNA helicase / DNA recombination / DNA repair / ATP hydrolysis activity / ATP binding / cytoplasm 類似検索 - 分子機能 | ||||||
生物種 | Streptococcus thermophilus (ストレプトコッカス・サリバリウス 亜種 サーモフィラス) Salmonella typhimurium (サルモネラ菌) synthetic construct (人工物) | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.1 Å | ||||||
データ登録者 | Goessweiner-Mohr, N. / Fahrenkamp, D. / Wald, J. / Marlovits, T.C. | ||||||
引用 | ジャーナル: Nature / 年: 2022 タイトル: Mechanism of AAA+ ATPase-mediated RuvAB-Holliday junction branch migration. 著者: Jiri Wald / Dirk Fahrenkamp / Nikolaus Goessweiner-Mohr / Wolfgang Lugmayr / Luciano Ciccarelli / Oliver Vesper / Thomas C Marlovits / 要旨: The Holliday junction is a key intermediate formed during DNA recombination across all kingdoms of life. In bacteria, the Holliday junction is processed by two homo-hexameric AAA+ ATPase RuvB motors, ...The Holliday junction is a key intermediate formed during DNA recombination across all kingdoms of life. In bacteria, the Holliday junction is processed by two homo-hexameric AAA+ ATPase RuvB motors, which assemble together with the RuvA-Holliday junction complex to energize the strand-exchange reaction. Despite its importance for chromosome maintenance, the structure and mechanism by which this complex facilitates branch migration are unknown. Here, using time-resolved cryo-electron microscopy, we obtained structures of the ATP-hydrolysing RuvAB complex in seven distinct conformational states, captured during assembly and processing of a Holliday junction. Five structures together resolve the complete nucleotide cycle and reveal the spatiotemporal relationship between ATP hydrolysis, nucleotide exchange and context-specific conformational changes in RuvB. Coordinated motions in a converter formed by DNA-disengaged RuvB subunits stimulate hydrolysis and nucleotide exchange. Immobilization of the converter enables RuvB to convert the ATP-contained energy into a lever motion, which generates the pulling force driving the branch migration. We show that RuvB motors rotate together with the DNA substrate, which, together with a progressing nucleotide cycle, forms the mechanistic basis for DNA recombination by continuous branch migration. Together, our data decipher the molecular principles of homologous recombination by the RuvAB complex, elucidate discrete and sequential transition-state intermediates for chemo-mechanical coupling of hexameric AAA+ motors and provide a blueprint for the design of state-specific compounds targeting AAA+ motors. | ||||||
履歴 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 7pbq.cif.gz | 651.9 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb7pbq.ent.gz | 546.1 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 7pbq.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/pb/7pbq ftp://data.pdbj.org/pub/pdb/validation_reports/pb/7pbq | HTTPS FTP |
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-関連構造データ
関連構造データ | 13299MC 7pblC 7pbmC 7pbnC 7pboC 7pbpC 7pbrC 7pbsC 7pbtC 7pbuC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 (文献) |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
-Holliday junction ATP-dependent DNA helicase ... , 2種, 7分子 ABCDEFG
#1: タンパク質 | 分子量: 35447.508 Da / 分子数: 6 / 由来タイプ: 組換発現 由来: (組換発現) Streptococcus thermophilus (ストレプトコッカス・サリバリウス 亜種 サーモフィラス) 遺伝子: ruvB, CDA68_01670, STHERMO_2112 発現宿主: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (大腸菌) 参照: UniProt: A0A2U2MES7, DNA helicase #2: タンパク質・ペプチド | | 分子量: 5105.734 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Salmonella typhimurium (サルモネラ菌) 遺伝子: ruvA, A3104_04360, A3S30_12815, A3T81_10475, A3U32_08040, A3V03_15555, A3V89_15755, A3W57_12595, A3W75_09730, A3X15_16725, A3X55_18545, A3Y76_18960, A4N07_18675, A4O05_13120, A4O41_13995, ...遺伝子: ruvA, A3104_04360, A3S30_12815, A3T81_10475, A3U32_08040, A3V03_15555, A3V89_15755, A3W57_12595, A3W75_09730, A3X15_16725, A3X55_18545, A3Y76_18960, A4N07_18675, A4O05_13120, A4O41_13995, A4R48_03555, A6D61_19645, AAA76_15415, AAB27_10170, AAB79_14680, AAC35_19210, ADQ28_18925, AF497_06575, AGM99_19520, AHN93_19355, AKH62_05145, AL144_07245, AL168_13275, AL184_08575, AQ530_20325, AU613_21305, AVA38_10310, AVC05_12030, AVL16_03380, AWT30_04170, AXX99_19885, B1265_18390, B1398_11015, B1642_21005, B1P38_18570, B2E31_18735, B4V59_02030, B4W90_19900, B6362_14325, B7Q27_17415, B8Y16_02315, B8Z46_03380, B9C90_13190, B9C96_10980, B9M14_03980, B9O84_20430, BBQ66_17875, BIC00_17935, BIC13_17075, BK110_18430, BKM50_18425, BMS46_18075, BMU56_19625, BZ203_15840, BZZ88_19070, C5W43_03605, CB102_15235, CB119_08750, CB198_07280, CB380_21505, CB535_05310, CB570_17460, CB646_18010, CBM67_10595, CBM76_08005, CBZ90_18715, CC339_18485, CC403_17275, CC453_11190, CC652_13440, CC971_18400, CCP17_18835, CDZ72_17145, CE70_01895, CED07_17535, CEQ70_19855, CFF58_03605, CFF59_03410, CHN22_03380, CIX60_20020, CPS79_06290, CQO33_10550, CSG22_19025, CVR97_18235, D4361_14120, D4387_19750, D4422_17805, D5823_07645, D5N86_14490, D5N95_15345, D5O82_08895, D5P17_20720, D5X47_14425, D5Y28_10680, D6422_17850, D6J79_19635, D8S24_16890, DD95_03300, DLB93_17215, DLR28_12360, DMI89_11085, DMO92_20225, DN165_16600, DNB97_11455, DNM27_19505, DNZ37_14615, DO533_14050, DP680_14360, DPB42_24680, DPD91_17340, DPF41_06595, DPF68_18980, DPS76_09640, DQD22_15400, DQR44_15500, DRM14_19730, DRR75_15340, DRT38_23380, DRT61_20665, DRV05_09715, DSF94_18945, DSG41_09900, DTF68_19605, DU071_21865, DU223_09180, DU657_10120, DU879_19615, DUV75_00760, DWU22_13690, DY580_20770, DYM27_19865, E0935_12270, E1A11_17010, E6W45_13400, EBD14_15340, EBK21_19660, EC404_05120, EEQ30_12650, EER35_11585, EHB09_02315, EL822_20025, ELS01_11305, EPB30_16090, EQG93_20735, EVY71_15105, EW905_19450, F0D96_03330, F2P00_04425, F3Q97_16890, F3R12_16560, F9G02_08090, F9O44_06330, FE758_08635, FEM52_20865, FGZ46_12135, FJM64_21435, FQC24_03995, FQX78_16495, G0038_05370, G0040_14630, G0042_05160, G0045_04645, G0047_04425, G0048_04645, G0051_04495, G0052_04480, G0059_16270, G0061_18365, G0062_04425, G0063_04645, G0067_04490, G0069_04495, G0070_04425, G0071_02335, G0072_18060, G0074_03845, G0076_09830, G0077_04570, G0080_04495, G0084_04425, G0086_04645, G0087_04645, G0088_03845, G0089_04305, G0090_18565, G0094_06110, G0100_04645, G0101_04930, G0102_18480, G0111_04570, G0113_07905, G0117_04640, G0123_04645, G0124_04480, G0148_04570, G0157_04060, G0170_04570, G0A05_20370, G0A28_08020, G0A32_09325, G0A39_13460, G0A43_07515, G0A44_14720, G0A46_17450, G0A50_13995, G0A51_15070, G0A52_16995, G0A53_13715, G0A56_10210, G0A58_13370, G0A60_08630, G0A61_09285, G0A63_17830, G0A66_14765, G0A67_09855, G0A68_07315, G0A70_13970, G0A73_14720, G0A76_13550, G0A79_10870, G0A85_06680, G0A88_03615, G0A92_16395, G0A96_15350, G0A97_09860, G0A99_04350, G0B03_14510, G0B05_11195, G0B07_12920, G0B08_11100, G0B12_13635, G0B28_07790, G0B96_17725, G0C03_09525, G0C04_00320, G0C34_16405, G0E15_19520, G0E20_18325, G0G84_09765, G0J24_00315, G0J26_18625, G0J27_17935, G0J28_18455, G0J31_17115, G0J33_11885, G0J34_00315, G0J36_20340, G0J37_00315, G0J40_15725, G0J43_14890, G0J44_19710, G0J45_19615, G0J46_20750, G0J47_00315, G0J49_00315, G0J50_19580, G0J51_19850, G0J53_15365, G0J55_18355, G0J58_16205, G0J59_19730, G0J62_01215, G0J65_19090, G0J66_17410, G0J67_02710, G0J69_19340, G0J71_17230, G0J73_20930, G0J76_18935, G0J79_15475, G0J81_04550, G0J82_17330, G0J85_04550, G0J89_20125, G0J92_18930, G0J94_20435, G0J96_20985, G0J97_19790, G0K00_14690, G0K02_19370, G0K03_16735, G0K04_20155, G0K05_08015, G0K07_19190, G0K10_18840, G0K13_14165, G0K15_20415, G0K16_18570, G0K18_18175, G0K19_15605, G0K20_08360, G0K23_16195, G0K25_19520, G0K26_18910, G0K28_18855, G0K30_19060, G0K31_17750, G0K32_07100, G0K33_18050, G0K37_17000, G0K38_19665, G0K39_12955, G0K41_06410, G0K42_19730, G0K44_18600, G0K46_04550, G0K47_00315, G0K48_19680, G0K49_19460, G0K52_20110, G0K53_20285, G0K56_14585, G0K58_16320, G0K59_20915, G0K61_12175, G0K65_17590, G0K68_19555, G0K70_19195, G0K72_08220, G0K74_09545, G0K75_14735, G0K78_04550, G0K80_18310, G0K83_17415, G0K84_18585, G0K85_19440, G0K88_000200, G0K89_000185, G0K90_000162, G0K94_000188, G0K95_000393, G0L00_001272, G0L02_000369, G0L03_03395, G0L06_03355, G0L07_06370, G0L10_06010, G0L14_04495, G0L15_00175, G0L18_02610, G0L19_04265, G0L20_06050, G0L24_11105, G0L25_13880, G0L29_13070, G0L31_13365, G0L32_11360, G0L34_10210, G0L35_10295, G0L36_12260, G0L37_14800, G0L38_03935, G0L40_08420, G0L42_07850, G0L48_05605, G0L49_04895, G0L51_16900, G0L52_12995, G0L55_17090, G0L59_12270, G0L62_12990, G0L63_12675, G0L65_16685, G0L67_15510, G0L68_16430, G0L70_19000, G0L73_10360, G0L76_16040, G0L77_17705, G0L78_16570, G0L79_13070, G0L83_12585, G0L85_15215, G0L86_003407, G0L88_08035, G0L89_12955, G0L91_13450, G0L93_14905, G0L96_17395, G0L98_14825, G0M00_15155, G0M02_15475, G0M05_07430, G0M06_002948, G0M10_06795, G0M13_002511, G0M14_09485, G0M16_09065, G0M18_001838, G0M21_12630, G0M22_002392, G0M25_002320, G0M26_13590, G0M27_18365, G0M28_14500, G0M29_002555, G0M30_14605, G0M33_08695, G0M35_17455, G0M36_13345, G0M38_11600, G0M39_14605, G0M40_09455, G0M41_12175, G0M45_15325, G0M46_002381, G0M48_002696, G0M51_10065, G0M53_11890, G0M55_12775, G0M56_13160, G0M58_13240, G0M63_12835, G0M65_12830, G0M67_12515, G0N45_03845, G0N48_13640, G0N51_02015, G0N53_12220, G0N55_03845, G0N57_13985, G0N58_03520, G0N59_04430, G0N60_03940, G0N61_04425, G0N62_02335, G0N64_03100, G0N65_20220, G0N66_03920, G0N67_14845, G0N71_17705, G0N75_04425, G0N78_15235, G0N82_20135, G0N84_04425, G0N85_21475, G0N86_03845, G0N88_00310, G0N89_04490, G0N90_19470, G0N92_13355, G0N94_13260, G0N95_12725, G0N98_18690, G0N99_19505, G0O00_17175, G0O03_13255, G0O10_03990, G0O14_11750, G0O15_19700, G0O18_03490, G0O19_04930, G0O20_08065, G0O22_16685, G0O25_17080, G0O27_03845, G0O31_03845, G0O32_09420, G0O36_02935, G0O37_17135, G0O39_04425, G0O40_04425, G0O41_16585, G0O42_18080, G0O43_04495, G0O44_02930, G0O47_03845, G0O52_03915, G0O55_03560, G0O57_17310, G0O58_04495, G0O59_04990, G0O60_03845, G0O63_03835, G0O66_19795, G0O68_03990, G0O70_04495, G0O71_04495, G0O74_19245, G0O75_04495, G0O77_04420, G0O78_03850, G0O80_03845, G0O81_18455, G0O82_04565, G0O84_04495, G0O85_04625, G0O86_20270, G0O87_02405, G0O88_03010, G0O89_04865, G0O92_04565, G0O93_11855, G0O94_19860, G0O97_21615, G0O99_16570, G0P00_02015, G0P01_14125, G0P02_18125, G0P05_20035, G0P06_04565, G0P08_18610, G0P12_02030, G0P13_04495, G0P17_18155, G0P18_20530, G0P19_11030, G0P24_14340, G0P26_04495, G0P28_04565, G0P30_04425, G0P31_04495, G0P36_10425, G0P37_21480, G0P41_15675, G0P44_15565, G0P45_15940, G0P48_04495, G0P49_12000, G0P52_04495, G0P53_12915, G0P56_03845, G0P57_04495, G0P58_03845, G0P63_02240, G0P65_03730, G0P67_18480, G0P68_10080, G0P69_06340, G0P73_15995, G0P75_14890, G0P76_22875, G1N61_16005, G1N64_15925, G1N66_15920, G1N68_15615, G1N71_15905, G1N72_15920, G1N86_16200, G1N87_15945, G1N91_16925, G1O00_16210, G1O02_16210, G1O04_15925, G1O05_15920, G1O08_17675, G1O10_10745, G1O12_16215, G1O16_16210, G1O17_15695, G1O18_16215, G1O20_16205, G1O23_15245, G1O25_16200, G1O26_16205, G1O27_16005, G1O28_15965, G1O29_15985, G1O32_16195, G1O34_16275, G1O38_16615, G1O40_16210, G1O43_16195, G1O46_15905, G1O48_15980, G1O49_16055, G1O51_16200, G1O53_16270, G1O62_16210, G1O63_16190, G1O65_15980, G1O67_15525, G1O68_16210, G1O69_15765, G1O71_15760, G1O72_16280, G1O76_16285, G1O77_16200, G1O80_16295, G1O81_15930, G1O83_16790, G1O84_16285, G1O87_16290, G1O88_16275, G1O89_16210, G1O90_16280, G1O93_16280, G1O94_16275, G1O96_16280, G1P02_16270, G1P03_17125, G1P06_15950, G1P09_16175, G1P10_16225, G1P12_16280, G1P14_14670, G1P15_16740, G1P17_16290, G1P19_16280, G1P23_16285, G1P24_16200, G1P25_17765, G1P26_16200, G1P29_17110, G1P31_16270, G1P35_16205, G1P36_16200, G1P37_17055, G1P40_16275, G1P44_16205, G1P45_16285, G1P47_16000, G1P48_16285, G1P51_16285, G1P52_17055, G1P53_17000, G1P54_16295, G1P55_15975, G1P56_16260, G1P57_16010, G1P58_16275, G1P59_16285, G1P61_15020, G1P64_15975, G1P67_16275, G1P69_16195, G1P72_12865, G1P75_15530, G1P76_15870, G1P78_19040, G1P83_18150, G1P84_16210, G1P87_16280, G1P90_21160, G1P91_16275, G1Q03_15890, G1Q08_16110, G1Q67_16285, G1Q78_14435, G1Q81_15765, G1Q83_16705, G1Q84_13315, G1Q85_15410, G1Q86_15380, G1Q88_15185, G1Q90_15690, G1Q91_15935, G1Q93_14530, G1Q96_15910, G1Q98_15430, G1Q99_11585, G1R01_17130, G1R02_16210, G1R03_16275, G1R04_16220, G1R08_13440, G1R13_15440, G1R15_13765, G1R20_17000, G1R21_15000, G1R22_15960, G1R23_13795, G1R27_15530, G1R28_15725, G1R29_16290, G1R30_15980, G1R31_15765, G1R36_17220, G1R38_14700, G1R40_15810, G1R42_16465, G1R44_16900, G1R45_15115, G1R47_15505, G1R48_13140, G1R51_15915, G1R52_13725, G1R53_17220, G1R63_16285, G1R69_13750, G1R87_16410, G1R93_15885, G1S02_16000, G2203_04570, G2212_12800, G2218_16175, G2221_09750, G2279_18630, G2290_14370, G2793_06395, G2918_19985, G2951_12275, G3221_002782, G3230_003711, G3231_003749, G3247_004406, G3248_003599, G3254_002478, G3263_003866, G3270_003535, G3275_002809, G3312_003088, G3336_004410, G3357_003961, G3369_002611, G3433_004280, G3460_003233, G3464_002156, G3593_002164, G3A35_17745, G3V06_002183, G3V14_004002, G3V17_003067, G3V21_003509, G3V56_002760, G3V57_004253, G3X03_002022, G4189_001963, G4190_002081, G4192_001775, G4198_002082, G4201_003454, G4202_003501, G4A01_004164, G4A73_002546, G4A83_001993, G4A85_003926, G4A87_002824, G4B68_002372, G4B72_002617, G4B74_002882, G4C74_003033, G4D32_000606, G4D46_001658, G4F88_13855, G4F89_10185, G4F91_09845, G4F92_14185, G4G47_003967, G4G62_001511, G4G67_002717, G4G68_002791, G4G75_002761, G4G76_002705, G4G79_003827, G4G97_002596, G4H00_003504, G4H04_001947, G4H07_003183, G4H08_003105, G4H18_002082, G4H21_003380, G4H24_003550, G4H63_003212, G4I66_003129, G4J07_003742, G4J08_004887, G4J11_001114, G4J12_002149, G4J18_000635, G4J20_003985, G4J37_003448, G4J39_003265, G4J41_000446, G4J45_002836, G4J90_003265, G4K02_003015, G4K03_002122, G4O54_002196, G4O56_001176, G4O59_002242, G4O60_002083, G4O67_002842, G4O69_000062, G4P29_004139, G4P83_003605, G4P85_003558, G4P89_003099, G4P91_003170, G4P93_003182, G4Q12_003157, G4Q28_002813, G4Q31_003329, G4Q50_001749, G4Q52_001957, G4Q59_003578, G4Q60_003633, G4Q63_004176, G4Q67_002937, G4Q94_002160, G4R01_003504, G4R02_003669, G4R15_002080, G4R16_003122, G4W68_002287, G4W73_002067, G4W86_002681, G4W87_003507, G4W88_003344, G4W91_003086, G4Y10_000342, G9269_003303, G9302_003971, G9304_003425, G9305_003122, G9309_001865, G9313_002530, G9314_002446, G9367_003273, G9381_002894, G9C24_004023, G9C41_001900, G9C46_002876, G9C47_001859, G9C49_002192, G9C57_003668, G9C64_003704, G9G03_002459, G9G04_000697, G9G34_003490, G9G36_002899, G9G45_003700, G9G50_002140, G9G62_004102, G9W19_004088, G9W28_003744, G9W45_001593, G9W52_003809, G9W63_003831, G9W65_003419, G9W79_003403, G9W95_002945, G9W96_002938, G9X40_003683, GB021_04425, GB040_15420, GB055_16785, GB076_20415, GB106_17280, GB114_20125, GB120_04410, GB122_20510, GB131_13210, GB139_16080, GB171_17175, GB209_03045, GB221_18125, GB224_14215, GB238_12855, GB280_04495, GB321_04495, GB331_20715, GB339_09370, GB342_19550, GB368_14700, GB372_17490, GB416_15465, GB452_14590, GB459_20995, GB466_20710, GB505_20505, GB510_11225, GB551_03055, GB567_19780, GB645_17180, GBS44_09110, GBS58_08305, GBV53_12830, GBV54_19265, GBV60_17435, GBW03_16075, GBW44_11995, GBW52_16250, GBW76_03040, GBX12_12465, GBX20_21570, GBX46_15100, GBX55_11165, GBX64_18390, GBY13_17555, GBY23_02020, GBY73_19470, GBZ51_23005, GBZ55_11025, GCZ80_03970, GEZ01_20335, GJE27_04470, GJE28_16295, GNA59_000161, GNA88_003884, GNA97_004062, GNA99_004226, GNB28_003525, GNB36_003216, GNB86_003069, GNB92_001265, GNC11_002998, GNC19_001983, GNC45_003243, GNC75_003299, GNC95_002454, GT380_17915, GTH60_10565, GTH62_16360, GTH63_03995, GTH66_01790, GTH67_13230, GTH68_05260, GTH70_11300, GTH72_02930, GTH73_04425, GTH75_04500, GTH77_15795, GTH78_16995, GTH79_16050, GTH81_13965, GTH85_04425, GTH87_03845, GTH89_13210, GTH90_11240, GTH91_20750, GTH93_04495, GTH94_02805, GTH99_12385, GXC51_17360, GXC56_17270, GXG40_17095, GYI58_17910, GYI62_002360, GYI77_20460, GYJ04_12525, GYJ24_04505, GYJ27_13680, GYJ28_001987, GYJ30_04425, GYJ32_04570, GYJ53_16290, GYJ59_13110, GYJ60_16265, H8S97_09740, KP44_16705, NG06_22005, R035_13700, SE14_02039, STMLT2P22_CBEKMEGD_02701, Z700_11380, ZV33_15625, ZX03_07070, ZY40_16825 発現宿主: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (大腸菌) 参照: UniProt: A0A0M0QTS9, DNA helicase |
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-DNA鎖 , 2種, 2分子 UV
#3: DNA鎖 | 分子量: 4643.037 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
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#4: DNA鎖 | 分子量: 4535.946 Da / 分子数: 1 / 由来タイプ: 合成 / 由来: (合成) synthetic construct (人工物) |
-非ポリマー , 3種, 10分子
#5: 化合物 | ChemComp-AGS / #6: 化合物 | ChemComp-MG / #7: 化合物 | |
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-詳細
研究の焦点であるリガンドがあるか | N |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 | 名称: RuvAB branch migration motor complexed to the Holliday junction - RuvB AAA+ state s0+A [t2 dataset] タイプ: COMPLEX / 詳細: RuvB helicase / Entity ID: #1-#4 / 由来: RECOMBINANT |
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分子量 | 値: 0.220 MDa / 実験値: NO |
由来(天然) | 生物種: Streptococcus thermophilus (ストレプトコッカス・サリバリウス 亜種 サーモフィラス) |
由来(組換発現) | 生物種: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (大腸菌) プラスミド: pProEX HTB |
緩衝液 | pH: 8 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: in-vitro reconstituted freshly before vitrification |
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil R2/2 |
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE-PROPANE / 湿度: 100 % / 凍結前の試料温度: 277 K |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 4000 nm / 最小 デフォーカス(公称値): 500 nm / Cs: 2.7 mm / C2レンズ絞り径: 70 µm / アライメント法: COMA FREE |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 5 sec. / 電子線照射量: 30.7 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 QUANTUM (4k x 4k) 実像数: 30083 |
電子光学装置 | エネルギーフィルター名称: GIF Bioquantum / エネルギーフィルタースリット幅: 10 eV |
-解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.1 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 62542 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: OTHER |